Biotinylated Human Siglec-2, Fc Tag, Avi Tag (SI2-H82F8) is expressed from human 293 cells (HEK293). It contains AA Asp 20 - Arg 687 (Accession # P20273-1).
Predicted N-terminus: Asp 20
This protein carries a human IgG1 Fc tag at the C-terminus, followed by a Avi tag (Avitag™).
The protein has a calculated MW of 103.9 kDa. The protein migrates as 115-120 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>95% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in 50 mM Tris, 100 mM Glycine, pH7.5. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human Siglec-2, Fc Tag, Avi Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Immobilized Anti-Human CD22 MAb at 0.5 μg/mL (100 μL/well) can bind Biotinylated Human Siglec-2, Fc Tag, Avi Tag (Cat. No. SI2-H82F8) with a linear range of 1-16 ng/mL (QC tested).
CAR阳性表达率检测（Evaluation of CAR expression ）
FACS Analysis of Anti-CD22 CAR Expression
293 cells were transfected with anti-CD22-scFv and RFP tag. 2e5 of the cells were first stained with B. Biotinylated Human Siglec-2 / CD22 Protein, Fc Tag, Avi Tag (Cat. No. SI2-H82F8, 10 μg/ml) and C. Biotinylated Protein Control, followed by FITC Streptavidin. A. Non-transfected 293 cells and C. Biotinylated Protein Control were used as negative control. RFP was used to evaluate CAR (anti-CD22-scFv) expression and FITC was used to evaluate the binding activity of Biotinylated Human Siglec-2 / CD22 Protein, Fc Tag, Avi Tag (Cat. No. SI2-H82F8).
FACS Analysis of Anti-CD22 CAR Expression
Human T cells were lentivirally transduced with anti-CD19 CAR and cultured for 3 days. Three days post-transduction, 1e6 cells were first incubated with 100ul Biotinylated Human Siglec-2 / CD22 Protein, Fc Tag, Avi Tag (Cat. No. SI2-H82F8, 10 μg/ml), washed and then stained with PE Streptavidin. Non-transduced T cells were used as a control for gating of CAR expression. (Data are kindly provided by Cellyan Therapeutics Co. Ltd.)
B-cell receptor CD22 is also known as Sialic acid-binding Ig-like lectin 2 (Siglec-2), B-lymphocyte cell adhesion molecule (BL-CAM), T-cell surface antigen Leu-14, which belongs to the immunoglobulin superfamily and SIGLEC (sialic acid binding Ig-like lectin) family. CD22 mediates B-cell B-cell interactions, and may be involved in the localization of B-cells in lymphoid tissues. Siglec-2 / CD22 binds sialylated glycoproteins, one of which is CD45. Siglec2 / CD22 plays a role in positive regulation through interaction with Src family tyrosine kinases and may also act as an inhibitory receptor by recruiting cytoplasmic phosphatases via their SH2 domains that block signal transduction through dephosphorylation of signaling molecules.
专利使用说明（Limited Use License）
The Biotin AviTag technology is covered by U.S. Pat. No: 5,874,239 and includes any and all materials, methods, kits and related derivatives claimed by this patent. The purchase of the Acrobiosystems’s Avitag™ proteins confers to the purchaser the limited right to use the Avitag™ technology for non-commercial, or research use, or for purposes of evaluating the Avitag™ technology.
Commercial use of the Avitag™ technology to manufacture a commercial product, or use of the Avitag™ technology to facilitate or advance research which will be applied to the development of a commercial product requires a license from Avidity, LLC. Examples of Commercial use include, but are not limited to biosensors, diagnostics, therapeutic applications, proximity assays, and drug screening assays.